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Chinese Livestock and Poultry Breeding ›› 2026, Vol. 22 ›› Issue (9): 103-112.doi: 10.19543/j.cnki.1673-4556.20260727.005cstr: 32418.14.j.cnki.1673-4556.20260727.005

• Regulation of Embryonic Development and Stem Cell Differentiation • Previous Articles     Next Articles

Research progress on the differentiation of domestic animal pluripotent stem cells into male germ cells

Qing Zhao(), Zhiyan Zhao, Xueling Li()   

  1. State Key Laboratory for Reproductive Regulation and Breeding of Grassland Livestock, Inner Mongolia University, Hohhot, 010020, Inner Mongolia
  • Received:2026-01-14 Online:2026-09-26 Published:2026-09-10
  • Contact: Xueling Li

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Abstract:

The traditional livestock breeding model relies on natural reproduction and phenotypic selection, which has limitations such as long generation interval, slow genetic progress, and difficulty in utilizing the excellent genetic material of sterile or dead individuals. In recent years, with the in-depth study of stem cell biology and epigenetic regulation mechanism, the use of pluripotent stem cells (PSCs) to reconstruct the gametogenesis process in vitro has become one of the breakthrough frontiers in the field of animal genetics and breeding. At present, the strategies for promoting the differentiation of livestock PSCs into male germ cells can be classified into three categories: step-by-step induction of cytokines, co-culture and microenvironment simulation, and directional induction of key genes. Although these strategies have been verified in marker detection and morphology, they still face problems such as low differentiation efficiency, high cell heterogeneity, and insufficient functional maturity of the obtained cells. Livestock PSCs have obvious species specificity, and the culture systems commonly used in mice and human cells cannot be directly applied to species such as pigs, cattle, and sheep. The maintenance of its pluripotency depends on the combination of different signaling pathways, and the initial cell state is different, which makes it difficult to universalize the differentiation programs between different species. At present, there is still a lack of a recognized standardized process. In addition, challenges such as technical complexity and ethical disputes also restrict its large-scale application. The future breakthrough direction can be focused on: developing customized differentiation medium according to the specific signal regulation map of species, so as to get rid of the limitation of empirical trial and error; at the same time, combined with microfluidic and organoid chip technology, the dynamic physical signals are coordinated with the regulation of biochemical factors, thereby improving the accuracy of the differentiation process. This paper systematically summarizes the research progress of the above three strategies in livestock, analyzes the key problems currently faced, and puts forward the future breakthrough direction, in order to provide theoretical reference for the follow-up research.

Key words: Livestock, Pluripotent stem cells, Male germ cells, Directional differentiation

CLC Number: 

  • S81

Fig. 1

Schematic diagram of the differentiation trajectory from livestock pluripotent stem cells into male germ cell lineages"

Table 1

Comparison of the principles and characteristics of three induction strategies"

策略

Strategies

原理

Principle

关键因子/技术

Key factors/Techniques

优势

Advantages

局限

Limitations

细胞因子分步诱导

Stepwise induction by cytokines

 模拟发育信号通路，时序性调控 BMP4、RA、CHIR99021等 理论基础明确，方案相对标准化 分化效率低，难以推进至功能性精子阶段

共培养与微环境模拟

Co-culture and microenvironment simulation

 重建生殖细胞-体细胞互作与空间结构

与支持细胞共培养；

3D类器官/水凝胶

 提供关键旁分泌信号，促进功能成熟

体系复杂、标准化难、

成本高

转入基因法

Transgenic methods

 直接操控命运决定基因

过表达SOX17等；

敲除负调控基因

 精准、高效，可绕过复杂信号 技术门槛高，可能引入基因组风险

Table 2

The representative results and verification of three induction strategies in livestock"

策略Strategies 应用示例Application examples 关键标志物Key markers 验证水平Verification level

细胞因子分步诱导

Stepwise induction by cytokines

猪PGCLCs诱导[30]

牛减数分裂启动[37]

 PRDM1、PRDM14、STELLA、DAZL、DDX4、SYCP3、REC8

基因表达：RT-qPCR/RNA-seq

蛋白表达：免疫荧光/Western blot

减数分裂启动：染色体铺展观察SYCP3丝状结构

共培养与微环境模拟

Co-culture and microenvironment simulation

牛PGCLCs高效诱导与分选[47]

猪SSCs与支持细胞共培养[43]

 DDX4、DAZL、SYCP3、γH2AX、GATA4、SOX9

基因表达：RT-qPCR/RNA-seq

蛋白表达：免疫荧光共定位

减数分裂结构：染色体铺展

功能互作：SSCs集落形成

转入基因法

Transgenic methods

 猪EPSCs高效诱导PGCLCs[15] PRDM1、SOX17、TFAP2C、NANOS3、DAZL、NANOG、OCT4

基因表达：RT-qPCR/RNA-seq

蛋白表达：免疫荧光/Western blot

发育潜能：嵌合体形成

功能验证：体内移植后生殖系贡献

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